Quantitative studies on the development of inoculated Brugia malayi microfilariae in Anopheles sinensis and Culex quinquefasciatus.

Definite numbers of B. malayi mf were inoculated into An. sinensis and Cx. quinquefasciatus, and their subsequent development was observed. The relationships between the dosage and the L3 positive rate on the one hand and the average filarial maturity rate on the other were defined. At the same dosages of 4 and 10 mf/mosquito, the L3 positive rates and the average filarial maturity rates in An. sinensis were much higher than those in Cx. quinquefasciatus (p less than 0.01). It was demonstrated that the immune response to filariae of the mosquitos was the main factor resulting in these differences. The intensity of the immune response not only varied with the species, but also declined with the aging of mosquitos. The results of our experiment might be useful to studies on the dynamics of filariasis transmission and the interaction between filariae and their insect hosts.

Comparative susceptibility of five strains of Mansonia uniformis (Diptera:Culicidae) in Malaysia to infection with subperiodic Brugia malayi.

Five strains of Ma. uniformis from Malaysia were tested for their susceptibility to infection with subperiodic B. malayi. All were found to be susceptible with infection rates ranging from 62% to 100%. The susceptibility rates were directly related to the microfilarial densities of the cat at the time of feeding. Statistical analysis showed no significant difference (p greater than 0.05) among the means of the indices of experimental infection as well as the percentage of infective mosquitoes of the five strains and an old laboratory colony. They were all equally susceptible to subperiodic B. malayi.

Cryopreservation of infective-stage larvae of Brugia malayi.

Third-stage larvae of Brugia malayi (Guizhou, China strain), in RPMI-1640, 6% DMSO and 15% calf serum, frozen slowly in the vapor phase of liquid nitrogen prior to emersion in liquid nitrogen, remained viable for as long as as 321 days and were infective to Mongolian jirds. All DMSO should be removed from the freezing medium to restore larval activity after freezing.

Development of Brugia malayi in Mongolian gerbils previously exposed to Wuchereria bancrofti.

Twelve Mongolian gerbils, Meriones unguiculatus, were infected with 100 third-stage larvae of Wuchereria bancrofti. One month later these animals, along with 4 control animals, were given 100 third-stage larvae of Brugia malayi. Eleven of the 12 experimental animals and the 4 controls survived, and 8 of the experimental animals and all of the controls demonstrated microfilaremia after 3 months. The animals were killed at 6-months post-infection and examined for parasites. One W. bancrofti larva was found in one of the experimental animals, and 15% of the B. malayi given were recovered as adults from the testes, viscera, and carcass. Thirty-eight percent of the worms given to the controls were recovered from the testes, viscera, and pelt. The worms from the experimental animals also appeared to be smaller. This study suggests that gerbils are able to develop partial resistance to Brugia malayi following a previous infection with Wuchereria bancrofti.

Brugia malayi: serum dependent cell-mediated reactions to microfilariae.

Sheathed and exsheathed microfilariae of Brugia malayi are killed by normal rat cells in the presence of immune serum in vitro. Immune serum heated at 56 degrees C for 1 hour lost this activity which was largely restored by the addition of fresh normal rat serum. EDTA but not EGTA abolished this activity indicating the operation of complement by alternate pathway. Fresh normal rat serum alone promoted cellular adherence without exerting cytotoxicity to the microfilariae. The activity in the immune serum could be removed with Staphylococcus aureus cells containing Protein A or anti-IgG antiserum. The activity could also be absorbed to and eluted from Protein A--sepharose CL-4B suggesting the involvement of IgG. Neutrophils and macrophages participate in the antibody dependent cell-mediated cytotoxicity phenomenon. Eosinophils while adhering to the microfilariae exert cytotoxicity only to the exsheathed parasites.

Studies on Malayan filariasis in Bengkulu (Sumatera), Indonesia with special reference to vector confirmation.

In an endemic focus of Brugia malayi in Bengkulu, Indonesia the microfilariae rate was 25%. The microfilariae showed nocturnal periodicity. Domestic cats in the same area were found to harbour B. pahangi, with a microfilariae rate of 23%. In the study area, the most prevalent man-biting Mansonia mosquitoes were Mn.bonneae (41%), followed by Mn.annulata (27%), Mn.uniformis (25%) and Mn.dives (7%). More Mansonia mosquitoes were collected outdoors than indoors between 1800-2200 hours. Natural infections with infective larvae of Brugia spp. were found in Mn.bonneae, Mn.dives, Mn.uniformis, Mn.annulata, and An.nigerrimus. Experimental infection studies revealed that Mn.annulata, Mn.bonneae, Mn.uniformis, An.nigerrimus and An.peditaeniatus permit the development of infective larvae. Under experimental condition, the vectorial competence is high in Mn.bonneae, intermediate in Mn.uniformis and low in An.hyrcanus group. It is concluded that the principal vectors of periodic B.malayi in the study area in Bengkulu are four Mansonia species (Mn.annulata, Mn.bonneae, Mn.uniformis and Mn.dives), and that the potential vectors include two Anopheles species (An.nigerrimus and An.peditaeniatus). This is the first record of Mn.bonneae being a vector of periodic B.malayi in Indonesia.

Mastomys natalensis as an experimental host for Brugia malaye subperiodic.

Infective larvae of Brugia malayi subperiodic obtained by dissection of infected Aedes togoi were injected subcutaneously into the scrotal region of Mastomys natalensis. From altogether 58 infected male M. natalensis 81% showed consistently or intermittently detectable microfilaraemia, whereas in 19% of the animals no microfilaraemia could be detected at any stage. The mean prepatent period was 136 days; the microfilarial density varied from 1 to 535 per 20 c. mm blood. In those animlas with consistently detectable and in general higher microfilaraemia an average of 13.1 live adult worms were found, against an average of 6.4 adult worms in animals with intermittent detectable and in general lower microfilaraemia. An average of 1.5 worms was found in animals which at no stage showed detectable microfilaraemia. A correlation between worm burden and prepatent period could be observed in the individual groups. From the total of 520 live adult worms recovered at necropsy, 37% were found in the lungs, 29% in the parenchyma of the testes and 34% in the lymphatic system. 47% of live fertile female worms were found in the lymphatic system, whereas the majority, i.e; 52% of infertile female worms were detected in the lungs. In addition, 380 encapsulated dead worms were found, most of them (98%) in the lymphatic system. 61% of a total of 900 live and dead worms were found in the region of the lymphatic system.

Experimental infection of Rattus sabanus and Rattus muelleri with subperiodic Brugia malayi.

R. sabanus and R. muelleri are very common in the lowland forests of Malaysia. In nature they are infected with Breinlia sp. and D. ramachandrani. In an attempt to determine whether they are also susceptible to subperiodic B. malayi and thereby being potential reservoirs of infection of the disease, 24 R. muelleri and 17 R. sabanus were experimentally infected with the parasite. Results show that although they can support the full development of the parasite, they are poor hosts. This confirms the observation that in Malaysia natural infection of Rattus spp. with the parasite has not been seen. These rats therefore are probably not important in the zoonotic transmission of subperiodic B. malayi in Malaysia.